Anti-Nestin antibody [2C1.3A11] (ab18102)

Product name

Anti-Nestin antibody [2C1.3A11]
See all Nestin primary antibodies
Description

Mouse monoclonal [2C1.3A11] to Nestin
Host species

Mouse
Tested applications

Suitable for: ICC/IF, WB, Flow Cyt, IHC-P, IPmore details
Species reactivity

Reacts with: Human
Does not react with: Rat
Immunogen

A 150 amino acid fragment from the cloned human Nestin.
General notes

This highly specific antibody to human nestin can aid in characterizing progenitor cells differentiating into distinct lineages, in enhancing the therapeutic potential of human neural stem/progenitor cells in the treatment of CNS diseases or injury, and in identifying neuroepithelial tumor cells.

Form

Liquid
Storage instructions

Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
Storage buffer

Preservative: 0.1% Sodium azide
Constituent: PBS
Concentration information loading...
Purity

Protein G purified
Primary antibody notes

This highly specific antibody to human nestin can aid in characterizing progenitor cells differentiating into distinct lineages, in enhancing the therapeutic potential of human neural stem/progenitor cells in the treatment of CNS diseases or injury, and in identifying neuroepithelial tumor cells.
Clonality

Monoclonal
Clone number

2C1.3A11
Isotype

IgG1
Research areas

Alternative Versions
- Anti-Nestin antibody [2C1.3A11] (HRP) (ab187845)
- Anti-Nestin antibody [2C1.3A11] (FITC) (ab187846)
Compatible Secondaries
Conjugation kits
- PE / R-Phycoerythrin Conjugation Kit (ab102918)
- APC Conjugation Kit (ab201807)
Isotype control
- Mouse IgG1, kappa monoclonal [15-6E10A7] - Isotype Control (ab170190)
- Mouse IgG1, Kappa Monoclonal [B11/6] - Isotype Control (ab91353)

Our Abpromise guarantee covers the use of ab18102 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application	Abreviews	Notes
ICC/IF		1/100 - 1/500.
WB		1/1000 - 1/5000. Predicted molecular weight: 177 kDa.
Flow Cyt		1/200. ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.
IHC-P		Use at an assay dependent concentration.
IP		Use at an assay dependent concentration.

Function

Required for brain and eye development. Promotes the disassembly of phosphorylated vimentin intermediate filaments (IF) during mitosis and may play a role in the trafficking and distribution of IF proteins and other cellular factors to daughter cells during progenitor cell division. Required for survival, renewal and mitogen-stimulated proliferation of neural progenitor cells.
Tissue specificity

CNS stem cells.
Sequence similarities

Belongs to the intermediate filament family.
Developmental stage

Upon terminal neural differentiation, nestin is down-regulated and replaced by neurofilaments.
Post-translational
modifications

Constitutively phosphorylated. This increases during mitosis when the cytoplasmic intermediate filament network is reorganized.
Target information above from: UniProt accession P48681 The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010) .

Information by UniProt
Database links
- Entrez Gene: 10763 Human
- Omim: 600915 Human
- SwissProt: P48681 Human
- Unigene: 527971 Human
Alternative names
- ESTM 46 antibody
- FLJ 21841 antibody
- FLJ21841 antibody
- Intermediate filament protein antibody
- Nbla00170 antibody
- nes antibody
- NEST_HUMAN antibody
- Nestin antibody
see all

Immunocytochemistry/ Immunofluorescence - Anti-Nestin antibody [2C1.3A11] (ab18102)

Immunofluorescence staining of U251 cells (glioblastoma cell line) stained using ab18102(green) and bisbenzimide (blue)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Nestin antibody [2C1.3A11] (ab18102)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of migrating tumor cells labelling Nestin with ab18102 at 2µg/ml. Cytoplasm and membrane positive staining shown.
Western blot - Anti-Nestin antibody [2C1.3A11] (ab18102)

All lanes : Anti-Nestin antibody [2C1.3A11] (ab18102) at 1/1000 dilution

Lane 1 : Total rat brain protein at 5 µg
Lane 2 : Human CNS progenitor cell protein at 1 µg
Lane 3 : Human CNS progenitor cell protein at 5 µg

Predicted band size: 177 kDa
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Nestin antibody [2C1.3A11] (ab18102)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of xenograft tumor cells labelling Nestin with ab18102.
Flow Cytometry - Anti-Nestin antibody [2C1.3A11] (ab18102)

Overlay histogram showing SH-SY5Y cells stained with ab18102 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab18102, 1/200 dilution) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x10⁶ cells) used under the same conditions. Acquisition of >5,000 events was performed.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Nestin antibody [2C1.3A11] (ab18102)

Immunohistochemistry of Anti-Nestin on formalin fixed paraffin embedded prostate nerve tissue using ab18102

Flow cytometry protocols
Immunoprecipitation protocols
Immunohistochemistry protocols
Immunocytochemistry & immunofluorescence protocols
Western blot protocols

This product has been referenced in:

Ferrer CM et al. An inactivating mutation in the histone deacetylase SIRT6 causes human perinatal lethality. Genes Dev 32:373-388 (2018). Read more (PubMed: 29555651) »
Jaraíz-Rodríguez M et al. A Short Region of Connexin43 Reduces Human Glioma Stem Cell Migration, Invasion, and Survival through Src, PTEN, and FAK. Stem Cell Reports 9:451-463 (2017). Read more (PubMed: 28712848) »

See all 4 Publications for this product

Publishing research using ab18102? Please let us know so that we can cite the reference in this datasheet.

Customer reviews and Q&As

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1-2 of 2 Abreviews or Q&A

Abcam guarantees this product to work in the species/application used in this Abreview.

Application

Flow Cytometry

Sample

Human Cell (Neural Progenitor cells (from human ES cells))

Specification

Neural Progenitor cells (from human ES cells)

Preparation

Cell harvesting/tissue preparation method: Cell Dissociation Buffer
Sample buffer: FACS buffer (0.2%BSA, 0.1%sodium azide in PBS)

Fixation

Paraformaldehyde

Permeabilization

Yes - 70% Methanol (ice cold)

Gating Strategy

FSC vs SSC on negative control sample

Abcam user community

Verified customer

Submitted Jan 03 2012

Question

Yes, that would be for the nestin antibodies only. (ab6320 & ab18102) If you can find any specific FACS protocol for one of these, then it may be decisive in my choice. Up to know, I do not have any information for choosing between them. Thus I am waiting for your answer. Anyway, if you cannot find any protocols,then I will choose one of them by chance. Thanks in advance, I hope this helps

Abcam community

Verified customer

Asked on Jun 18 2007

Answer

I am please to be able to share the following protocol for ab18102for Flow cytometry: Fix cells with 4% Paraformaldehyde for 30 min at room temperature (RT) Wash twice with 1% PBS at 1500rpm/5min/RT Permeabilize cells with 0.02% Triton X-100 in 1%PBS + 1% BSA for 15 min/RT Then add rabbit nestin ab (1:400 dilution) for 1hr/RT Wash twice Add specific secondary ab for 1hr/RT Wash twice Flow reading I hope this information will be helpful. Please do not hesitate to contact me if I can be of further assistance,

Abcam Scientific Support

Answered on Jun 18 2007

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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Anti-Nestin antibody [2C1.3A11] (ab18102)

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Flow Cytometry abreview for Anti-Nestin antibody [2C1.3A11] - Neural Stem Cell Marker